Prokaryotic expression vector construction,expression,purification and identification of human protein tyrosine kinase 6

We aimed to prepare purification PTK6 recombinant protein for antibody preparation and functional analysis.Firstly,we used PCR for adding EcoR I and BamH I enzyme sites in the ORF of PTK6.After purification and enzyme digestion,the fragment were subcloned into prokaryotic expression vector PHUE to construct recombinant protein expression vector PHUE/PTK6,which identified by PCR,enzyme digestion and sequence analysis,and then transform into E.coli BL21(DE3).Then IPTG was used to induce the recombinant protein expression.SDS-PAGE analysis suggested the recombinant protein was expressed as inclusion body,with molecular weight of about 55 000,which consistent with the expecting value.After washing,the recombinant protein was purified by affinity chromatography and the purity coefficient was up to 80%.Purified human PTK6 recombinant protein prepared in this study will facilitate antibody preparation,structure investigation and protein functional analysis.